Leflunomide active metabolite inhibites the expression of phorbol-12-myristate-13-acetate-induced CD147,matrix metallo-proteinase-2 and matrix metallo-proteinase-9 on THP-1 cells
10.3760/cma.j.issn.1007-7480.2011.03.011
- VernacularTitle:来氟米特活性代谢产物抑制人单核细胞系细胞CD147基质金属蛋白酶-2及基质金属蛋白酶-9表达的研究
- Author:
Shiyao WU
;
Jianlin HUANG
;
Baozhao XIE
;
Mingxia WANG
;
Jieruo GU
- Publication Type:Journal Article
- Keywords:
Gelatinase A;
Gelatinase B;
Antigens,CD147
- From:
Chinese Journal of Rheumatology
2011;15(3):183-187,后插1
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of the leflunomide active metabolite (A771726) on the expression of phorbol-12-myristate-13-acetate (PMA) -induced CD147, matrix metallo-proteinase (MMP)-2 and MMP-9 on THP-1 cells. Methods THP-1 cells were cultured in RPMI-1640 medium supplemented with 10％ fetal bovine serum. For all experiments, THP-1 cells were cultured at an initial density of 5×105/ml. Before A771726 treatment, cells were cultured with serum-free RPMI-1640 medium for 12 h, THP-1cells were co-cultured with PMA at three different concentrations of A771726 (5, 15 , 45 μg/ml) for 24 h.The mRNA expression of CD147, MMP-2 and MMP-9 was measured by real-time PCR. CD147 expression on the cells were evaluated by flow cytometric analysis. The activity of MMP-2 and MMP-9 were evaluated by gelatin zymography. Statistical differences among the groups were tested by one-way ANOVA or KruskalWallis test. Results The expression of CD147, MMP-2 and MMP-9 were upgraded by the PMA. The expression of CD147 on THP-1 cells was inhibited significantly by A771726 in a dose-dependent pattern (P＜0.01). The mean fluorescence intensity (MFI) of CD147 in positive control group was 109.5±3.8, the MFI in A771726 (5, 15, 45 μg/ml) group were 73.3±2.5, 64.5±2.3, 40.9±2.7, respectively. The expression of MMP-2, MMP-9 mRNA and the activity of MMP-2, MMP-9 in the supernatant was inhibited significantly by A771726 (P＜0.01). The expression of CD147 mRNA was not inhibited significantly by A771726 (P＞0.05).Conclusion Leflunomide active metabolite (A771726) can inhibit the expression of PMA-induced CD147,MMP-2 and MMP-9 on THP-1 Cells.
